目的 探讨脑泰方是否通过调控热休克转录因子1（heat shock factor 1，HSF1）/热休克蛋白B1（heat shock proteins B1，HSPB1）通路降低脑铁沉积、抑制活性氧沉积及脂质过氧化产物产生、减轻神经元铁死亡，从而发挥脑卒中缺血损伤保护作用。方法 SD大鼠随机分为假手术组、模型组、脑泰方（27 g/kg）组和去铁酮（125 mg/kg）组，给予药物干预。通过大脑中动脉闭塞（middle cerebral artery occlusion，MCAO）法制备脑缺血大鼠模型，术后2 h进行神经功能评分及取材。TTC染色法检测脑梗死体积；苏木素-伊红（HE）染色观察脑组织形态结构及损伤情况；尼氏染色观察脑组织尼氏体数目；普鲁士蓝染色观察脑组织神经元内铁聚集水平；免疫组化及Western blotting法检测脑组织HSF1、HSPB1、转铁蛋白受体1（transferrin receptor 1，TFR1）和铁蛋白重链多肽（ferritin heavy polypeptide1，FTH1）蛋白表达；铁离子检测试剂盒与丙二醛检测试剂盒及活性氧检测试剂盒检测脑组织铁、丙二醛及活性氧含量。结果 与模型组比较，脑泰方组大鼠脑梗死体积减少，神经功能评分降低（P<0.01）；脑缺血区尼氏体增多，细胞损伤程度降低；含铁聚集颗粒细胞的数目减少；脑铁、丙二醛及活性氧含量降低（P<0.05、0.01）；脑组织HSF1、HSPB1及FTH1蛋白表达上调（P<0.05、0.01），TFR1蛋白表达下调（P<0.05）。结论 脑泰方可能通过上调HSF1/HSPB1通路，抑制TFR1的表达以减少神经元铁的吸收，上调FTH1的表达以增加铁蛋白的铁存储，以此调控铁代谢稳态平衡，进而抑制因过量的铁通过芬顿反应产生的活性氧及随后脂质过氧化产物引起的缺血性脑卒中神经元铁死亡。

Objective To explore whether Naotaifang (脑泰方) reduces brain iron deposition by regulating heat shock factor 1 (HSF1)/heat shock proteins B1 (HSPB1) pathway, inhibiting the deposition of reactive oxygen species and the production of lipid peroxidation products, reducing neuronal ferroptosis, and exerting a protective effect on cerebral ischemia injury. Methods SD rats were randomly divided into sham group, model group, Naotaifang (27 g/kg) group ang deferiprone (125 mg/kg) group, drugs were given for intervention. Cerebral ischemia rats model was prepared by middle cerebral artery occlusion (MCAO) method, and the neurological function was scored 2 h after operation. TTC staining was used to detect cerebral infarct volume; HE staining was used to observe the morphology and damage; Nissl staining was used to observe the number of Nissl bodies; Prussian blue staining was used to observe the level of iron accumulation in neurons; Western blotting and immunohistochemical experiments werre used to detect the protein expression levels of HSF1, HSPB1, transferrin receptor 1 (TFR1) and ferritin heavy polypeptide1 (FTH1); Iron ion detection kits, malondialdehyde detection kits and reactive oxygen detection kits were used to detect brain iron, malondialdehyde and reactive oxygen content. Results Compared with model group, the cerebral infarction volume of rats in Naotaifang group was decreased, and neurological function score was decreased (P < 0.01); Nissl bodies in cerebral ischemia area was increased, and the degree of cell damage was decreased; Number of iron-containing aggregate granule cells were decreased; Brain iron, malondioxide aldehyde and reactive oxygen content were decreased (P < 0.05, 0.01); HSF1, HSPB1 and FTH1 protein expressions were up-regulated (P < 0.05, 0.01), and TFR1 protein expression was down-regulated (P < 0.05). Conclusion Naotaifang may up-regulate HSF1/HSPB1 pathway, inhibit the expression of TFR1 to reduce neuronal iron absorption, and increase the expression of FTH1 to increase the iron storage of ferritin, thereby regulating the steady-state balance of iron metabolism, inhibiting excessive ferroptosis of neurons in ischemic stroke caused by active oxygen produced by Fenton reaction and subsequent lipid peroxide.

R285.5

湖南省自然科学基金面上项目（2020JJ4467）；湖南省教育厅科学研究重点项目（19A378）；湖南省教育厅科学研究项目（18C0379）